A cDNA library of the venom of Inimicus japonicus was constructed. The cDNA was cloned into eukaryotical expression plasmid pcDNA3.0. SMART^TM protocol was used for cDNA library construction and bioinformatics analysis was carried out. 94 novel EST clones were obtained from 150 sequences in the library, of which there were 35 full-length clones, including cytolysin genes, short neurotoxin gene, C-type lectin gene, macrophage migration inhibitory factor gene and so on. Most of those genes were reported for the first time in Inimicus japonicus. Further studies on those genes and large scales of sequencing in the library are going on, which may be helpful to make clear the components of Inimicus japonicus venom and understand the function of those proteins at molecular level.